CN106822993B - Nerve graft and nerve graft system using same - Google Patents
Nerve graft and nerve graft system using same Download PDFInfo
- Publication number
- CN106822993B CN106822993B CN201710211602.5A CN201710211602A CN106822993B CN 106822993 B CN106822993 B CN 106822993B CN 201710211602 A CN201710211602 A CN 201710211602A CN 106822993 B CN106822993 B CN 106822993B
- Authority
- CN
- China
- Prior art keywords
- nerve
- bundle channel
- channel
- sensory
- motor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/227—Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
- A61L27/3675—Nerve tissue, e.g. brain, spinal cord, nerves, dura mater
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Transplantation (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Medicinal Chemistry (AREA)
- Dermatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Neurology (AREA)
- Vascular Medicine (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Neurosurgery (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Dispersion Chemistry (AREA)
- Prostheses (AREA)
Abstract
The invention provides a nerve graft and a nerve graft system using the same, and relates to the technical field of medical engineering. The nerve graft provided by the invention comprises a sensory nerve bundle channel, a motor nerve bundle channel and a mixed nerve bundle channel, wherein each nerve bundle channel is matched with a nerve bundle of a corresponding segment of a normal nerve and is respectively filled with NGF, GDNF, NGF and GDNF. The nerve graft provided by the invention is matched with the ganglion segment to be repaired in appearance, and the shape of the channel of the inner nerve bundle is also accurately matched with the repaired nerve bundle. And secondly, different neurotrophic factors are loaded in the nerve graft respectively, so that the nerve bundles with different functions can be promoted and guided in directions, the matching rate of the nerve graft and the receptor nerve can be improved, the classified guidance of different nerve bundles is realized, and the nerve repair effect is improved. In addition, the invention also provides a nerve graft system applying the nerve graft to treat complex nerve defect patients.
Description
Technical Field
The invention relates to the technical field of medical engineering, in particular to a nerve graft and a nerve graft system using the same.
Background
At present, the clinical gold standard for repairing peripheral nerve defects is autologous nerve transplantation, namely, other nerves with relatively insignificant functions are sacrificed to be used as a bridge body for repairing the nerves with relatively important functions, and a better repairing effect can be usually achieved. Other potential nerve grafts include both nerve conduits and decellularized allogenic nerves. The nerve conduit is an empty conduit prepared from degradable natural or synthetic materials, can guide and support the regeneration of peripheral nerves, prevent the lateral growth of the nerves, limit the growth of myofibroblasts, reduce the formation of scars and the like. Another class of products is decellularized allogeneic nerves. Sondel et al first reported in 1998 that chemical extraction was used to decellularize rat sciatic nerves, removing cells and debris from nerve tissue, and obtaining a decellularized nerve scaffold composed of extracellular matrix (ECM). Since the acellular nerve scaffold retains the structure of the natural nerve, it is also effective for large nerve defects as long as 5 cm. The university of maryland jia dao peak team reports for the first time that a Y-shaped nerve conduit loaded with different neurotrophic factors is prepared by computer-aided modeling design and 3D printing technology in 2015, so that the classified guidance of motor nerves and sensory nerves is realized.
However, the autologous nerves have limited sources, are all fine sensory nerves, have poor matching with the nerves to be repaired, cause permanent nerve dysfunction of donor areas, traumatic neuroma, skin scar and the like, and are not ideal nerve repair schemes. The nerve conduit is mainly used for sleeving and protecting a nerve suture opening or bridging a small nerve short segment defect (the nerve defect of not more than 3 cm) at present, and for acellular allogeneic nerves, donor nerves matched with damaged nerves need to be selected for accurately repairing the nerve defect, are limited by a human tissue donor source, and the cell-free nerve scaffold customized according to the repairing requirement is high in difficulty, high in cost and low in efficiency. Furthermore, most of the currently used nerve grafts do not distinguish between sensory nerves, motor nerves and mixed nerves, but only a tubular nerve graft is singly arranged and filled with the same filler indiscriminately, as shown in fig. 1, and the sensory nerves, the motor nerves and the mixed nerves cannot be treated or transplanted respectively in a targeted manner.
Therefore, how to improve the bionic degree of the structure and the microenvironment in the nerve graft, highly match the sensory nerve, the motor nerve and the mixed nerve at the defect part and reduce the mismatch rate of nerve regeneration has important significance.
Disclosure of Invention
The invention aims to provide a nerve graft so as to relieve the technical problem of low bionic degree of the structure and the microenvironment of the nerve graft in the prior art.
The invention provides a nerve graft, which comprises a sensory nerve fasciculus channel, a motor nerve fasciculus channel and a mixed nerve fasciculus channel, wherein the distribution and the walking shape of the sensory nerve fasciculus channel, the motor nerve fasciculus channel and the mixed nerve fasciculus channel are matched with the distribution and the walking shape of a nerve fasciculus of a corresponding section of a normal nerve;
the sensory nerve bundle channel comprises a sensory nerve bundle channel tube, and a sensory nerve bundle channel filler filled in the sensory nerve bundle channel tube, the sensory nerve bundle channel filler comprising NGF;
the motor nerve bundle channel comprises a motor nerve bundle channel tube, and a motor nerve bundle channel filler filled in the motor nerve bundle channel tube, the motor nerve bundle channel filler comprising GDNF;
the mixed nerve bundle channel comprises a mixed nerve bundle channel tube, and a mixed nerve bundle channel filler filled in the mixed nerve bundle channel tube, the mixed nerve bundle channel filler comprising NGF and GDNF.
Further, the distribution and the shape-walking regular data of the sensory nerve tract channel, the motor nerve tract channel and the mixed nerve tract channel are derived from a three-dimensional visual digital model of the nerve tracts of the corresponding segments of the normal nerve.
Further, the wall thickness of the sensory nerve bundle passage tube, the motor nerve bundle passage tube and the mixed nerve bundle passage tube is 0.05-0.6mm.
Further, the sensory nerve bundle channel filler, the motor nerve bundle channel filler and the mixed nerve bundle channel filler each further comprise a hydrogel; the nerve graft as claimed in claim 3, wherein the hydrogel is prepared by peripheral nerve decellularization.
Further, in the above-mentioned case,
in the sensory nerve bundle channel filler, the concentration of the NGF is 50-100ng/mL;
in the motor nerve bundle channel filler, the concentration of GDNF is 50-100ng/mL;
in the mixed nerve bundle channel filler, the concentration of the NGF is 50-100ng/mL, and the concentration of the GDNF is 50-100ng/mL.
Further, in the above-mentioned case,
the sensory nerve bundle channel further comprises a sensory nerve bundle channel support disposed inside the sensory nerve bundle channel tube;
the motor nerve bundle channel also comprises a motor nerve bundle channel bracket arranged inside the motor nerve bundle channel tube;
the hybrid nerve bundle channel further comprises a hybrid nerve bundle channel stent disposed inside the hybrid nerve bundle channel tube.
Further, the sensory nerve bundle channel tube, the motor nerve bundle channel tube, the mixed nerve bundle channel tube, the sensory nerve bundle channel scaffold, the motor nerve bundle channel scaffold and the mixed nerve bundle channel scaffold are made of a biocompatible degradable material.
Further, the biocompatible degradable material is PLA and/or chitosan.
Further, the nerve graft is prepared by a 3D printing method.
In addition, the invention also provides a nerve graft system, and the nerve graft system applies one or more nerve grafts.
The nerve graft provided by the invention is matched with the ganglion segment to be repaired in appearance, and the distribution and the walking shape of the inner nerve bundle channel are also matched with the repaired nerve bundle accurately. Secondly, different nerve bundle channels in the nerve graft are loaded with different neurotrophic factors, and can promote and guide the nerve bundles with different functions. The nerve graft provided by the invention can improve the matching rate of the nerve graft and the receptor nerve, realize the classification guidance of sensory nerve tracts, motor nerve tracts and mixed nerve tracts and improve the nerve repair effect.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a prior art nerve graft;
fig. 2 is a schematic structural diagram of a nerve graft provided in example 1 of the present invention;
FIG. 3 is a schematic structural view of a nerve graft provided in example 2 of the present invention;
fig. 4 is a schematic cross-sectional view of a motor nerve bundle channel provided in example 2 of the present invention.
Icon: 1-sensory nerve bundle channel; 11-sensory nerve bundle channel tube; 12-sensory nerve bundle channel filler; 13-sensory nerve bundle channel scaffold; 2-motor nerve bundle channel; 21-motor nerve bundle channel tube; 22-motor tract channel filler; 23-motor tract channel scaffold; 3-mixed nerve bundle channels; 31-mixed nerve bundle channel tube; 32-mixed nerve bundle channel filler; 33-hybrid nerve bundle channel scaffolds.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and it is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a nerve graft, which comprises a sensory nerve fasciculus channel 1, a motor nerve fasciculus channel 2 and a mixed nerve fasciculus channel 3, wherein the distribution and the walking shape of the sensory nerve fasciculus channel 1, the motor nerve fasciculus channel 2 and the mixed nerve fasciculus channel 3 are matched with the distribution and the walking shape of nerve fasciculus of corresponding segments of normal nerves; wherein the sensory nerve bundle channel 1 comprises a sensory nerve bundle channel tube 11, and a sensory nerve bundle channel filler 12 filled in the sensory nerve bundle channel tube 11, the sensory nerve bundle channel filler 12 comprising NGF; the motor nerve bundle channel 2 comprises a motor nerve bundle channel tube 21, and a motor nerve bundle channel filler 22 filled in the motor nerve bundle channel tube 21, the motor nerve bundle channel filler 22 comprising GDNF; the mixed bundle channel 3 includes a mixed bundle channel tube 31, and a mixed bundle channel filler 32 filled in the mixed bundle channel tube 31, the mixed bundle channel filler 32 including NGF and GDNF.
Wherein, NGF (NGF) and GDNF (GDNF) are both neurotrophic factors.
NGF has a selective chemotactic effect on the growth of sensory neuron axons, and the filling of NGF in the sensory nerve bundle channel tube 11 can further promote and induce the growth of the sensory nerve bundle channel 1 in the right direction; GDNF has selective chemotactic effect on the growth of motor neuron axons, and the filling of GDNF in the motor nerve bundle channel tube 21 can further promote and induce the growth of the motor nerve bundle channel 2 along the correct direction; filling the mixed nerve bundle channel tube 31 with NGF and GDNF further promotes and induces the growth of the mixed nerve bundle channel 3 in the right direction. By filling different neurotrophic factors, the microenvironment of the nerve graft provided by the invention is kept highly consistent with that of a normal nerve, and the high bionics of the microenvironment is embodied.
It is emphasized that the distribution and the shape-change rule data of the sensory nerve fasciculus channel 1, the motor nerve fasciculus channel 2 and the mixed nerve fasciculus channel 3 of the nerve graft provided by the invention are derived from a nerve fasciculus three-dimensional visualization digital model of a corresponding segment of a normal nerve. In the actual operation process, the normal nerve segment of the part to be repaired of the patient can be analyzed, and after three-dimensional data are collected, the shape, the structure and the trend of the whole nerve graft are determined according to the corresponding data. The nerve graft manufactured according to the method keeps the maximum consistency with the normal nerve section of a patient, embodies the high imitativeness of the structure, and has higher guiding and promoting effects on the repair of damaged nerves.
The nerve graft provided by the invention has no fixed limiting shape, and needs to be set according to the anatomical structure of the normal nerve segment of a part to be transplanted of a patient, namely, the specific corresponding shapes and relative positions of the sensory nerve tract channel 1, the motor nerve tract channel 2 and the mixed nerve tract channel 3 are set according to specific conditions.
The nerve graft provided by the invention is matched with the ganglion segment to be repaired in appearance, and the shapes of the sensory nerve bundle channel 1, the motor nerve bundle channel 2 and the mixed nerve bundle channel 3 are also highly matched with the repaired nerve bundle. Secondly, the sensory nerve bundle channel filler 12, the motor nerve bundle channel filler 22 and the mixed nerve bundle channel filler 32 in the nerve graft are respectively loaded with different neurotrophic factors, and can promote and guide the nerve bundles with different functions.
The sensory nerve bundle passage tube 11, the motor nerve bundle passage tube 21 and the mixed nerve bundle passage tube 31 of the nerve graft provided by the present invention are all made of biocompatible degradable materials, such as, but not limited to, polylactic acid (PLA), collagen fiber, polycaprolactone (PCL), polyglycolic acid (PGA), polylactic-glycolic acid copolymer (PGLA), dioxanone Polymer (PDS), chitosan, etc., preferably PLA and/or chitosan.
After the nerve graft provided by the invention is implanted into a nerve defect part, the sensory nerve bundle channel tube 11, the motor nerve bundle channel tube 21 and the mixed nerve bundle channel tube 31 of the nerve graft are gradually degraded, and the sensory nerve bundle channel filler 12, the motor nerve bundle channel filler 22 and the mixed nerve bundle channel filler 32 gradually complete the nerve regeneration process.
The wall thickness of the nerve graft channel tube 11, the motor tract channel tube 21 and the mixed nerve tract channel tube 31 provided by the present invention is 0.05-0.6mm, and may be, for example, 0.05mm, 0.06mm, 0.07mm, 0.08mm, 0.09mm, 0.1mm, 0.11mm, 0.12mm, 0.13mm, 0.14mm, 0.15mm, 0.16mm, 0.17mm, 0.18mm, 0.19mm, 0.2mm, 0.21mm, 0.22mm, 0.23mm, 0.24mm, 0.25mm, 0.26mm, 0.27mm, 0.28mm, 0.29mm, 0.3mm, 0.31mm, 0.32mm, 0.33mm, 0.34mm, 0.35mm, 0.36mm, 0.37mm, 0.38mm, 0.39mm, 0.3mm, 0.31mm, 0.32mm, 0.33mm, 0.34mm, 0.35mm, 0.36mm, 0.37mm, 0.54mm, 0.53mm, 0.48mm, 0.53mm, 0.48mm, or 49 mm.
The nerve graft provided by the invention has the advantages that the sensory nerve bundle channel filler 12, the motor nerve bundle channel filler 22 and the mixed nerve bundle channel filler 32 also comprise hydrogel, namely, the sensory nerve bundle channel filler 12 consists of the hydrogel and NGF, the motor nerve bundle channel filler 22 consists of the hydrogel and GDNF, and the mixed nerve bundle channel filler 32 consists of the hydrogel and the NGF and GDNF.
In the invention, the hydrogel is prepared by carrying out acellular manipulation on peripheral nerves, and specifically comprises the following steps:
taking fresh peripheral nerves, shearing off adipose tissues and partial nerve adventitia on the surface of the fresh peripheral nerves under an operation microscope, and placing the treated nerves in distilled water for shaking and rinsing for 6 hours;
step (b), extraction: placing nerves into 3% Triton X-100 water solution, shaking for 12h, rinsing in distilled water for 3 times, placing into 4% sodium deoxycholate water solution, shaking for 24h at room temperature, and rinsing in distilled water for 3 times; the extraction process is circulated twice;
step (c), lyophilization: putting the extracted nerves into a mixed solution of ethanol and dichloromethane for degreasing, wherein the volume ratio of the ethanol to the dichloromethane is 1;
and (d) dissolving and digesting: adding hydrochloric acid solution into the freeze-dried powder for dissolving to obtain an acellular matrix, adding pepsin into the acellular matrix to obtain a digestive juice, wherein the mass ratio of the pepsin to the acellular matrix is 1;
step (e), adjusting the pH value of the digestive juice to be more than 8 by using NaOH solution, and adjusting the pH value to be less than 7.4 to obtain a pre-gel solution; adding 10 XPBS to the neutral pre-gel solution at 4 ℃, wherein the volume ratio of the 10 XPBS to the pre-gel solution is 1;
and (f) culturing the pre-gel solution at 37 ℃ to obtain the acellular nerve hydrogel.
After the hydrogel is prepared according to this method, NGF, or GDNF, or NGF and GDNF, are dissolved in the hydrogel, i.e., the sensory, motor, and mixed fascial channel fillers 12, 22, and 32, respectively, are formed. Wherein the concentration of NGF in sensory nerve bundle channel filler 12 is 50-100ng/mL, for example 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100ng/mL; wherein the concentration of GDNF in motor tract channel filler 22 is 50-100ng/mL, for example, 50ng/mL, 55ng/mL, 60ng/mL, 65ng/mL, 70ng/mL, 75ng/mL, 80ng/mL, 85ng/mL, 90ng/mL, 95ng/mL, or 100ng/mL; wherein the concentration of NGF in mixed bundle channel filler 32 is 50-100ng/mL, for example 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100ng/mL; GDNF may be present in a concentration of 50-100ng/mL, for example, 50ng/mL, 55ng/mL, 60ng/mL, 65ng/mL, 70ng/mL, 75ng/mL, 80ng/mL, 85ng/mL, 90ng/mL, 95ng/mL, or 100ng/mL.
In addition, a stent can be arranged in three nerve tracts of the nerve graft provided by the invention, namely a sensory nerve tract channel stent 13 is also arranged in the sensory nerve tract channel 1, a motor nerve tract channel stent 23 is also arranged in the motor nerve tract channel 2, and a mixed nerve tract channel stent 33 is also arranged in the mixed nerve tract channel 3. The sensory nerve bundle channel stent 13, the motor nerve bundle channel stent 23 and the hybrid nerve bundle channel stent 33 are made of biocompatible degradable materials, such as, but not limited to, polylactic acid (PLA), collagen fiber, polycaprolactone (PCL), polyglycolic acid (PGA), poly (lactic-co-glycolic acid) (PGLA), dioxanone Polymer (PDS), chitosan, etc., preferably PLA and/or chitosan. Similarly, after the nerve graft provided by the present invention is implanted into a nerve defect site, the sensory tract channel scaffold 13, the motor tract channel scaffold 23 and the mixed tract channel scaffold 33 of the nerve graft are gradually degraded.
The number of the sensory nerve bundle channel stents 13 in the nerve graft provided by the invention is 10-60, for example, 10, 15, 20, 25, 30, 35, 40, 55 or 60; the sensory nerve bundle channel scaffold 13 has a diameter of 0.02-0.08mm, and may be, for example, 0.02mm, 0.025mm, 0.03mm, 0.035mm, 0.04mm, 0.045mm, 0.05mm, 0.055mm, 0.06mm, 0.065mm, 0.07mm, 0.075mm, or 0.08mm; the number of the motor nerve bundle channel brackets 23 is 10-60, for example, 10, 15, 20, 25, 30, 35, 40, 55 or 60; the motor nerve bundle channel support 23 has a diameter of 0.02-0.13mm, for example, may be 0.02mm, 0.025mm, 0.03mm, 0.035mm, 0.04mm, 0.045mm, 0.05mm, 0.055mm, 0.06mm, 0.065mm, 0.07mm, 0.075mm, 0.08mm, 0.09mm, 0.10mm, 0.11mm, 0.12mm, or 0.13 mm; the number of the mixed nerve bundle channel brackets 33 is 10-60, for example, 10, 15, 20, 25, 30, 35, 40, 55 or 60; the diameter of the mixed nerve bundle channel scaffold 33 is 0.02-0.08mm, and may be, for example, 0.02mm, 0.025mm, 0.03mm, 0.035mm, 0.04mm, 0.045mm, 0.05mm, 0.055mm, 0.06mm, 0.065mm, 0.07mm, 0.075mm, or 0.08mm.
The nerve graft provided by the invention is prepared by a 3D printing method, and different areas are printed by different 3D printing inks.
In addition, the invention also provides a nerve graft system, and different specifications of the nerve graft provided by the invention are determined according to different specific diseased conditions of patients.
In order to facilitate a clearer understanding of the disclosure of the present invention, reference will now be made in detail to specific embodiments thereof.
Example 1
Fig. 2 is a schematic structural diagram of the nerve graft provided in this embodiment, and as shown in fig. 2, the sensory nerve bundle passage 1 and the motor nerve bundle passage 2 meet to form a mixed nerve bundle passage 3, forming a "Y" shaped structure, in this embodiment, the wall thicknesses of the sensory nerve bundle passage tube 11, the motor nerve bundle passage tube 21 and the mixed nerve bundle passage tube 31 are all 0.23mm. Sensory neurofascial channel filling 12 consists of hydrogel and NGF, wherein the concentration of NGF is 85ng/mL; the motor tract channel filler 22 consists of hydrogel and GDNF, wherein the concentration of GDNF is 80ng/mL; the mixed bundle channel filler 32 consists of hydrogel and NGF and GDNF at concentrations of 85ng/mL and 80ng/mL, respectively.
The preparation method of the hydrogel in the embodiment comprises the following steps:
taking peripheral nerves of a mouse, shearing off fat tissues and partial nerve adventitia on the surface of the peripheral nerves under a surgical microscope, and placing the treated nerves in distilled water for shaking and rinsing for 6 hours;
step (b), extraction: putting nerves into a Triton X-100 aqueous solution with the concentration of 3% for oscillation for 12h, then rinsing the nerves in distilled water for 3 times, then putting the nerves into a sodium deoxycholate aqueous solution with the concentration of 4% for oscillation for 24h at room temperature, and finally rinsing the nerves in distilled water for 3 times; the extraction process is circulated twice;
step (c), lyophilization: putting the extracted nerves into a mixed solution of ethanol and dichloromethane for degreasing, wherein the volume ratio of the ethanol to the dichloromethane is 1;
step (d), dissolution and digestion: adding hydrochloric acid solution into the freeze-dried powder for dissolving to obtain an acellular matrix, adding pepsin into the acellular matrix to obtain a digestive juice, wherein the mass ratio of the pepsin to the acellular matrix is 1;
step (e), adjusting the pH value of the digestive juice to be more than 8 by using NaOH solution, and adjusting the pH value to be less than 7.4 to obtain a pre-gel solution; adding 10 XPBS to the neutral pre-gel solution at 4 ℃, wherein the volume ratio of the 10 XPBS to the pre-gel solution is 1;
and (f) culturing the pre-gel solution at 37 ℃ to obtain the acellular nerve hydrogel.
After the hydrogel is prepared according to this method, NGF, or GDNF, or NGF and GDNF, are dissolved in the hydrogel, i.e., the sensory, motor, and mixed fascial channel fillers 12, 22, and 32, respectively, are formed.
The preparation method of the nerve graft in this example is as follows: preparing materials (chitosan) of a sensory nerve bundle channel tube 11, a motor nerve bundle channel tube 21 and a mixed nerve bundle channel tube 31, a sensory nerve bundle channel filler 12 (hydrogel with NGF concentration of 85 ng/mL), a motor nerve bundle channel filler 22 (hydrogel with GDNF concentration of 80 ng/mL), and a mixed nerve bundle channel filler 32 (hydrogel with NGF concentration of 85ng/mL and GDNF concentration of 80 ng/mL) in a cartridge of a 3D printer; adjusting settings of the printer; the nerve graft is prepared by accurately simulating the three-dimensional structure of the nerve of the region to be transplanted and performing high-degree simulation printing.
Example 2
Example 2 provides a nerve graft which is an improvement on example 1, compared with example 1, example 2 provides a nerve graft which is further provided with a sensory nerve bundle channel stent 13, a motor nerve bundle channel stent 23 and a hybrid nerve bundle channel stent 33, as shown in fig. 3 and 4, fig. 3 is a schematic structural diagram of the nerve graft provided in this example; fig. 4 is a schematic cross-sectional view of the motor nerve bundle channel 2 provided in the present embodiment; wherein fig. 3 and 4 only represent schematic diagrams of the nerve graft, and do not limit the number or diameter of the specific stent.
In the nerve graft provided by the embodiment, the sensory nerve bundle channel support 13, the motor nerve bundle channel support 23 and the mixed nerve bundle channel support 33 are respectively arranged in the sensory nerve bundle channel 1, the motor nerve bundle channel 2 and the mixed nerve bundle channel 3, wherein the number of the sensory nerve bundle channel support 13, the motor nerve bundle channel support 23 and the mixed nerve bundle channel support 33 is 15, the diameter is 0.08mm, and the nerve graft can further accelerate nerve regeneration of a damaged area besides a supporting function.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (8)
1. A nerve graft, characterized in that the nerve graft comprises a sensory nerve fascicle passage (1), a motor nerve fascicle passage (2) and a mixed nerve fascicle passage (3), and the distribution and the walk shape of the sensory nerve fascicle passage (1), the motor nerve fascicle passage (2) and the mixed nerve fascicle passage (3) are matched with the distribution and the walk shape of the nerve fascicle of the corresponding segment of a normal nerve;
the sensory nerve bundle channel (1) comprises a sensory nerve bundle channel tube (11), and a sensory nerve bundle channel filler (12) filled in the sensory nerve bundle channel tube (11), the sensory nerve bundle channel filler (12) comprising NGF;
the motor nerve bundle channel (2) comprises a motor nerve bundle channel tube (21), and a motor nerve bundle channel filler (22) filled in the motor nerve bundle channel tube (21), the motor nerve bundle channel filler (22) comprising GDNF;
the mixed bundle channel (3) comprises a mixed bundle channel tube (31), and a mixed bundle channel filler (32) filled in the mixed bundle channel tube (31), the mixed bundle channel filler (32) comprising NGF and GDNF;
the sensory nerve bundle channel filler (12), the motor nerve bundle channel filler (22), and the hybrid nerve bundle channel filler (32) each further comprise a hydrogel; the hydrogel is prepared by peripheral nerve decellularization;
the sensory nerve bundle channel (1) further comprises a sensory nerve bundle channel stent (13) disposed inside the sensory nerve bundle channel tube (11);
the motor nerve bundle channel (2) further comprises a motor nerve bundle channel bracket (23) arranged inside the motor nerve bundle channel tube (21);
the hybrid nerve bundle channel (3) further comprises a hybrid nerve bundle channel stent (33) disposed inside the hybrid nerve bundle channel tube (31);
the hydrogel is prepared by the transcellular operation of peripheral nerves, and comprises the following steps:
taking fresh peripheral nerves, shearing off adipose tissues and partial nerve adventitia on the surface of the fresh peripheral nerves under an operation microscope, and placing the treated nerves in distilled water for shaking and rinsing for 6 hours;
step (b), extraction: putting nerves into a Triton X-100 aqueous solution with the concentration of 3% for oscillation for 12h, then rinsing the nerves in distilled water for 3 times, then putting the nerves into a sodium deoxycholate aqueous solution with the concentration of 4% for oscillation for 24h at room temperature, and finally rinsing the nerves in distilled water for 3 times; the extraction process is circulated twice;
step (c), lyophilization: putting the extracted nerves into a mixed solution of ethanol and dichloromethane for degreasing, wherein the volume ratio of the ethanol to the dichloromethane is 1;
and (d) dissolving and digesting: adding hydrochloric acid solution into the freeze-dried powder for dissolving to obtain an acellular matrix, adding pepsin into the acellular matrix to obtain a digestive juice, wherein the mass ratio of the pepsin to the acellular matrix is 1;
step (e), adjusting the pH value of the digestive juice to be more than 8 by using NaOH solution, and adjusting the pH value to be less than 7.4 to obtain a pre-gel solution; adding 10 XPBS to the neutral pre-gel solution at 4 ℃, wherein the volume ratio of the 10 XPBS to the pre-gel solution is 1;
and (f) culturing the pre-gel solution at 37 ℃ to obtain the acellular nerve hydrogel.
2. The nerve graft as claimed in claim 1, characterised in that the regular data of the distribution and the course of the sensory tract channel (1), the motor tract channel (2) and the mixed tract channel (3) are derived from a three-dimensional visual digital model of the tract of the corresponding segment of the normal nerve.
3. The nerve graft as claimed in claim 2, wherein the wall thickness of the sensory-nerve-bundle passage tube (11), the motor-nerve-bundle passage tube (21) and the mixed-nerve-bundle passage tube (31) is 0.05-0.6mm.
4. The nerve graft of claim 1,
in the sensory nerve bundle channel filler (12), the concentration of the NGF is 50-100ng/mL;
(ii) in said motor nerve bundle channel filler (22), the concentration of said GDNF is 50-100ng/mL;
in the mixed bundle channel filler (32), the concentration of NGF is 50-100ng/mL and the concentration of GDNF is 50-100ng/mL.
5. The nerve graft according to claim 1, wherein the sensory nerve bundle channel tube (11), the motor nerve bundle channel tube (21), the mixed nerve bundle channel tube (31), the sensory nerve bundle channel scaffold (13), the motor nerve bundle channel scaffold (23) and the mixed nerve bundle channel scaffold (33) are made of biocompatible degradable material.
6. The nerve graft as claimed in claim 5, wherein the biocompatible degradable material is PLA and/or chitosan.
7. The nerve graft as claimed in claim 6, wherein the nerve graft is made by a 3D printing method.
8. A nerve graft system, wherein one or more of the nerve grafts of claim 7 are used.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211602.5A CN106822993B (en) | 2017-03-31 | 2017-03-31 | Nerve graft and nerve graft system using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211602.5A CN106822993B (en) | 2017-03-31 | 2017-03-31 | Nerve graft and nerve graft system using same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106822993A CN106822993A (en) | 2017-06-13 |
| CN106822993B true CN106822993B (en) | 2023-02-03 |
Family
ID=59141336
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710211602.5A Active CN106822993B (en) | 2017-03-31 | 2017-03-31 | Nerve graft and nerve graft system using same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106822993B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109550083B (en) * | 2018-11-21 | 2021-04-23 | 中山大学附属第一医院 | Plugging device for preventing traumatic neuroma formation and preparation method thereof |
| CN110227184B (en) * | 2019-07-16 | 2020-04-24 | 南通大学 | Differential tissue engineered nerves and applications |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1258591C (en) * | 2004-05-18 | 2006-06-07 | 中山大学附属第一医院 | Tissue engineered peripheral nerve graft |
| EP2296751A1 (en) * | 2008-06-03 | 2011-03-23 | Medtronic, Inc. | Bifurcated lead system and apparatus |
| CN207722144U (en) * | 2017-03-31 | 2018-08-14 | 中山大学附属第一医院 | Nerve graft and apply its nerve graft system |
| CN110585488B (en) * | 2019-10-29 | 2021-12-28 | 中国医科大学 | Nerve repair catheter prepared from novel composite material and preparation method thereof |
-
2017
- 2017-03-31 CN CN201710211602.5A patent/CN106822993B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN106822993A (en) | 2017-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20210030528A1 (en) | Medical/Surgical Implant | |
| US8858631B2 (en) | Synthetic scaffolds and organ and tissue transplantation | |
| KR20100046037A (en) | Prosthesis for promoting the in vivo reconstruction of a hollow organ or a portion of a hollow organ | |
| WO2015185219A1 (en) | Production of a component for cultivating a joint surface implant | |
| Colombo et al. | Regenerative medicine: clinical applications and future perspectives | |
| US10213296B2 (en) | Graft including osseous tissue and cartilage tissue for augmentation rhinoplasty, and method of performing augmentation rhinoplasty using the graft | |
| CN104490491A (en) | Biological-compatibility artificial ear and in-vitro rapid construction method thereof | |
| CN106822993B (en) | Nerve graft and nerve graft system using same | |
| CN104740686A (en) | Step-by-step tissue engineering bone building method | |
| TWI450735B (en) | Composition for repairing cartilage tissue and method for making the same | |
| CN207722144U (en) | Nerve graft and apply its nerve graft system | |
| Thygesen et al. | Comparison of off-the-shelf β-tricalcium phosphate implants with novel resorbable 3D printed implants in mandible ramus of pigs | |
| CN103394126B (en) | Cartilage repair material and preparation method thereof | |
| Llames et al. | Tissue bioengineering and artificial organs | |
| US20220257365A1 (en) | Implant for reconstruction of nipple-areola complex, and method for producing same | |
| CN104874024A (en) | Cell assembling small-intestinal submucosa bionic composite engineering bone and preparation method thereof | |
| Chiesa et al. | Three-dimensional and four-dimensional printing in otolaryngology | |
| CN211561236U (en) | Bone defect repair support | |
| CN114945391A (en) | 3D bioprinting scaffold for tissue regeneration | |
| Kale et al. | A novel perfusion-based method for cochlear implant electrode insertion | |
| CN206576988U (en) | A kind of plunger type porous support for eye socket bone defect healing | |
| Lai et al. | Combining Piezoelectric Stimulation and Extracellular Vesicles for Cartilage Regeneration | |
| Rizzi | Personalized valvular tissue engineering: a scaled up recellularization procedure to produce functional tissue for living aortic valve substitutes | |
| Dinaki et al. | Journal of Clinical and Translational Research | |
| Kade | 3D printed structures with biomimicry mechanical characteristics for auricular cartilage growth |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |